We have developed a novel model in which to study neurotoxicity associated with ethanol withdrawal. The model utilizes organotypic cultures of neonatal rat hippocampus with neurotoxicity assessed by propidium iodide uptake quantified by imaging under fluorescence microscopy. Alcohol withdrawal, achieved by changing the culture medium, is associated with significant regional neurotoxicity in these cultures, and this is at least partly a consequence of glutamate/NMDA receptor activation. The proposal is to evaluate the role of endogenous polyamines in the co-activation of these NMDA receptors. This will be achieved by a series of pharmacological investigations designed to block either the effects of polyamines on the NMDA receptor, or to reduce the synthesis of polyamines in the culture system. Finally, the hypothesis will be tested directly by measuring the content of different polyamines in the culture and medium during withdrawal of ethanol in vitro. The proposal is of direct relevance to mechanisms of alcohol withdrawal, and to potential novel treatments of alcohol-induced neurodegeneration.